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Cloning, expression and immunological evaluation of a short fragment from Rv3391 of Mycobacterium tuberculosis

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Abstract

Tuberculosis (TB) is still one of the greatest health care problems in the world. In order to identify antigens that may be used in the serodiagnosis of active tuberculosis, a short fragment from Rv3391 (fRv3391) of Mycobacterium tuberculosis was cloned and expressed. Its molecular weight and secondary structure elements were identified by mass spectrometry and circular dichroism. And its immunological nature was also evaluated by enzyme-linked immunosorbent assay (ELISA). The fRv3391 was expressed in Escherichia coli BL21 (DE3) with the molecular weight of 42.5 kDa and the secondary structure elements 36.2% α-helix, 0.0% β-sheet, 32.6% β-turn, and 31.3% random coil. Evaluation of fRv3391 as an ELISA solid-phase antigen on a set of human sera from well-characterized TB cases and healthy subjects revealed that there was strong serum antibody reactivity to fRv3391 in many human TB patients. Taken together, a short fragment from Rv3391 of Mycobacterium tuberculosis was cloned and expressed and the ELISA results showed that the protein may be useful as an immunodominant antigen for the serodiagnosis of active TB.

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Acknowledgments

This study was supported by China Mega-Projects of Science Research for the 11th Five Year Plan (No. 2008ZX10003-003-03) and Shanghai Commission of Science and Technology, PR China (No. 08410703800).

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Bioengineering, Zhengzhou University, Zhengzhou, 450001, Henan, China

    Jun-wei Zhao, Yan-feng Gao & Yuan-ming Qi

  2. Department of Microbiology and Parasitology, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200025, China

    Jun-wei Zhao, Zhan-qiang Sun, Xiao-kui Guo & Shu-lin Zhang

  3. Linyi Chest Hospital, Linyi, Shandong, China

    Hong-guo Yang

  4. Institute of Genetics, Fudan University, Shanghai, China

    Chong-zhen Zhang

  5. School of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, Hubei, China

    Xiao-li Yu & Zi-lu Wen

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  1. Jun-wei Zhao
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  2. Zhan-qiang Sun
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  3. Hong-guo Yang
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  4. Chong-zhen Zhang
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  5. Xiao-li Yu
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  7. Yan-feng Gao
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  8. Xiao-kui Guo
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  9. Yuan-ming Qi
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  10. Shu-lin Zhang
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Correspondence to Yuan-ming Qi or Shu-lin Zhang.

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Jun-wei Zhao and Zhan-qiang Sun contributed equally to this work.

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Zhao, Jw., Sun, Zq., Yang, Hg. et al. Cloning, expression and immunological evaluation of a short fragment from Rv3391 of Mycobacterium tuberculosis . Ann Microbiol 61, 345–353 (2011). https://doi.org/10.1007/s13213-010-0148-7

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  • DOI: https://doi.org/10.1007/s13213-010-0148-7

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